Basanta Kumar Das

Expression patterns and mutation analysis of p53 in fish Rita rita from polluted riverine environment

The present study was undertaken to investigate the alterations in gene expression patterns and for mutation analysis of p53 in the riverine catfish Rita rita collected from polluted riverine habitat. The partial p53 gene sequence of Rita rita generated showed a high degree of similarities with the DNA binding domains of fishes, mice and human. Transcriptomic analysis, carried out by quantitative real-time Polymerase Chain Reaction (RT-qPCR), showed significant down-regulation of p53 in fishes collected from most of the polluted stretches. Similar trend in protein abundance was observed by western blot analysis. Down-regulation of p53 was more pronounced in gill than liver. Expression patterns of p53 suggest that exposure to a multitude of contaminants in the natural riverine ecosystem could suppress the expression of p53. Genomic DNA showed a low stained smear pattern upon electrophoresis, with no evidence of DNA fragmentation. For mutation analysis PCR-SSCP followed by sequence analysis was carried out, which identified eight mutations; two at codon level and six missense mutations in the DNA binding domain IV and V. Secondary structure prediction showed that these mutations could lead to impairment of protein structure. Thus, the present study indicated that aquatic pollution has impacted these lower vertebrates which are reflected by the down-regulation of tumor suppressor protein (p53) in majority of the stretches studied.

The population structure and genetic divergence of Labeo gonius (Hamilton, 1822) analyzed through mitochondrial DNA cytochrome b gene for conservation in Indian waters

Abstract The present study explains the population structure and genetic diversity of medium carp Labeo gonius by analyzing partial sequence of mitochondrial DNA cytochrome b gene. Labeo gonius is a lower risk Near Threatened species, distributed throughout the North Indian major rivers, reservoirs and lakes. This species has a larger scope as an alternative candidate species in carp aquaculture system. In the present investigation, 223 individuals of Labeo gonius were collected from five locations of phylo-geographically isolated riverine ecosystems of India resulted in 12 haplotypes. These haplotypes showed 14 variables, out of which 9 were singletons and 5 were parsimony informative sites of nucleotide positions. The haplotypes H1 was considered as ancestral haplotype. All the haplotypes were connected to each other by 1–4 nucleotide mutations. The Narmada haplotypes (H10; H11 and H12) were isolated from H1 by four nucleotide mutations. The analyses resulted maximum expansion events (τ = 4.13672) in Narmada, with Fst scores more than other population pairs. The analysis of molecular variance (AMOVA) showed significant genetic differentiation among populations (ØST = 0.69470, p < .000). The genetic differentiation patterns were significantly consistence with geographical distributions. This study rejected the null hypothesis of single panmictic population of medium carp, Labeo gonius in Indian water.

Genetic variation in wild and hatchery population of Catla catla (Hamilton, 1822) analyzed through mtDNA cytochrome b region

Abstract Catla (Catla catla) is a one of the most harvested Indian major carps and is widely cultured fish species in Indian subcontinent. In the present study, genetic variability between hatchery and wild stocks of Catla was surveyed using sequence data of mitochondrial DNA of partial 307 bp of cytochrome b region. A total of 174 Catla individuals were examined from three different river basins and hatcheries. Significant genetic heterogeneity was observed for the sequence data (FST = 0.308, p ≤ 0.001). However, analysis of molecular variance (AMOVA) resulted in insignificant genetic differentiation among the samples of three rivers and culture zones (FCT = −0.10, p = 0.44). The result suggested a significant genetic variation within different riverine system, low genetic differentiation among samples from river basins and a lack of genetic variation in hatchery populations.

Environmental DNA (eDNA): A Promising Biological Survey Tool for Aquatic Species Detection

Aquatic species are facing at higher risk of extinction similar to that of any other living components of diversified ecosystem in present scenario. So that, the conservation of aquatic biodiversity is much more important to know about the accurate information regarding species composition and their biological community interactions. Generally, traditional survey methods depend on physical identification and characterization of species but it has some sorts of challenging chances due to the phenotypic plasticity, sibling species, different stages of life cycle and its invasiveness. To overcome such barriers one of the significant and promising tool likewise environmental DNA (eDNA), which way the collection of genetic materials from bulk environment (i.e. soil, water, sediment etc.) circuitously from organisms has been used to monitor and analyzed the biodiversity status, invasive species along with the species of conservation category. Recently, the real application of eDNA analysis based outcomes uphold the actual emerging know how practices in support of the population and community ecology, conservation biology as well as in the superior field of taxonomical research. Such scientific appraisal will be useful in understanding the brief history of aquatic eDNA and obviously its methodological considerations, gentle sources, collection and analysis process, physical form, its persistence and proper transport in aquatic ecosystem. Moreover, the fruitful drives for summarization the discoveries of eDNA application and method over traditional technique, its recent challenges and examine the current and future frontiers along with the appropriate practices of aquatic eDNA relevancy in aquatic ecosystem.

Deep insights into the mode of ATP-binding mechanism in Zebrafish cyclin-dependent protein kinase-like 1 (zCDKL1): A molecular dynamics approach

In eukaryotes, the serine/threonine kinases (STKs) belonging to cyclin-dependent protein kinases (CDKs) play significant role in control of cell division and curb transcription in response to several extra and intra-cellular signals indispensable for enzymatic activity. The zebrafish cyclin-dependent protein kinase-like 1 protein (zCDKL1) shares a high degree of sequence and structural similarity with mammalian orthologs and express in brain, ovary, testis, and low levels in other tissues. Regardless of its importance in the developmental process, the structure, function and mode of ATP recognition have not been investigated yet due to lack of experimental data. Henceforth, to gain atomistic insights in to the structural dynamics and mode of ATP binding, a series of computational techniques involving theoretical modeling, docking, molecular dynamics (MD) simulations and MM/PBSA binding free energies were employed. The modeled bi-lobed zCDKL1 shares a high degree of secondary structure topology with human orthologs where ATP prefers to lie in the central cavity of the bi-lobed catalytic domain enclosed by strong hydrogen bonding, electrostatic and hydrophobic contacts. Long range MD simulation portrayed that catalytic domain of zCDKL1 to be highly rigid in nature as compared to the complex (zCDKL1-ATP) form. Comparative analysis with its orthologs revealed that conserved amino acids i.e., Ile10, Gly11, Glu12, Val18, Arg31, Phe80, Glu 130, Cys143 and Asp144 were crucial for ATP binding mechanism, which needs further investigation for legitimacy. MM/PBSA method revealed that van der Waals, electrostatic and polar solvation energy mostly contributes towards negative free energy. The implications of ATP binding mechanism inferred through these structural bioinformatics approaches will help in understanding the catalytic mechanisms of important STKs in eukaryotic system.

Therapeutic applications of zebrafish (Danio rerio) miRNAs linked with human diseases: A prospective review

MicroRNAs (miRNAs) are the class of small, non-coding RNAs that are produced from precursor transcripts by subsequent processing steps mediated by members of the RNaseIII family, Dicer and Drosha protein within cell. The importance of zebrafish miRNAs in regulation of normal cellular development and support to various kinds of metabolism process. Although the zebrafish model provides a fundamental platform for the study of developmental biology but recent work with zebrafish model has expanded its appliance to a broad range of experimental studies relevant to different kind of human diseases. Presently, the zebrafish model is used for the study of cardiovascular disease, schizophrenia, bipolar I disorder in eyes, psoriasis, spinal cord injury, cancer and diabetes that showing in some selected miRNAs are regulate these diseases in molecular levels. Here, a superior drive performed to depict the fundamental utilization of the zebrafish miRNAs that targeted to several clinical diseases connected to human. This review aims to provide a summary of understanding of the cellular mechanism which is responsible for selected diseases and suggests some therapeutic application for inhibition of miRNA functions.

Nutrigenomic studies on hilsa to evaluate flesh quality attributes and genes associated with fatty acid metabolism from the rivers Hooghly and Padma

The Indian shad hilsa (Tenualosa ilisha), a commercially important food fish rich in oils, enjoys high consumer preference in the South Asian countries owing to its unique flavour and culinary properties. The present study was undertaken with the primary objective of determining the flesh quality attributes of hilsa in terms of nutritive value (gross chemical composition, amino acid, fatty acid and mineral composition), pH, water holding capacity (WHC) and expression of genes associated with fatty acid metabolism and flesh quality. Additionally, comparative studies on the flesh quality attributes in hilsa from two distributaries of river Ganga i.e. Hooghly and Padma were also carried out. A high WHC (>80%) suggested juicy and tender nature of hilsa meat. The protein content was 18-21% in hilsa from both the rivers and essential amino acid lysine, valine and functional amino acids leucine and arginine were significantly higher in Hooghly hilsa (P<0.05). The predominance of umami taste amino acids, glutamic acid and aspartic acid and sweet taste amino acids, serine, glycine and alanine in hilsa from both the rivers could be the contributing factors to its unique flavour. The fat content in hilsa from river Hooghly and Padma were found to be 9.94 and 7.84%, respectively. The concentration of flavouring fatty acids like saturated fatty acids (SFA) (myristic acid) and omega (ω)-3 polyunsaturated fatty acids (PUFAs) (linoleic acid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, EPA and docosahexaenoic acid, DHA) were significantly higher in Hooghly hilsa (P<0.05). Among the genes associated with fatty acid metabolism studied, expression of cluster of differentiation (CD36), acetyl CoA oxidase (ACO), fatty acid synthase (FAS), peroxisome proliferator activated receptor beta (PPARβ), peroxisome proliferator activated receptor gamma (PPARγ) and desaturase were significantly higher in Padma hilsa (P<0.05), and the change was <2 fold. Comparative gene expression profiling of flesh quality genes (actin, GAPDH, LDH, TPI) showed similar levels of expression in hilsa from both the rivers (P<0.05). The nutrigenomic information generated on various flesh quality attributes of hilsa has enriched the knowledgebase. Further, from comparative nutrient analysis on hilsa from river Hooghly and Padma, it was observed that Hooghly hilsa is superior in terms of oil content, ω-3 PUFAs EPA and DHA and essential amino acids; however, the expression profile of genes associated with flesh quality were found to be similar. Thus, within the scope of the present study, Hooghly hilsa (medium size category, 500-700g size) was found to be nutritionally superior.

Polycyclic Aromatic Hydrocarbons (PAHs) in inland aquatic ecosystems: Perils and remedies through biosensors and bioremediation

Polycyclic Aromatic Hydrocarbons (PAHs) are among the most ubiquitous environmental pollutants of high global concern. PAHs belong to a diverse family of hydrocarbons with over one hundred compounds known, each containing at least two aromatic rings in their structure. Due to hydrophobic nature, PAHs tend to accumulate in the aquatic sediments, leading to bioaccumulation and elevated concentrations over time. In addition to their well-manifested mutagenic and carcinogenic effects in humans, they pose severe detrimental effects to aquatic life. The high eco-toxicity of PAHs has attracted a number of reviews, each dealing specifically with individual aspects of this global pollutant. However, efficient management of PAHs warrants a holistic approach that combines a thorough understanding of their physico-chemical properties, modes of environmental distribution and bioaccumulation, efficient detection, and bioremediation strategies. Currently, there is a lack of a comprehensive study that amalgamates all these aspects together. The current review, for the first time, overcomes this constraint, through providing a high level comprehensive understanding of the complexities faced during PAH management, while also recommending future directions through potentially viable solutions. Importantly, effective management of PAHs strongly relies upon reliable detection tools, which are currently non-existent, or at the very best inefficient, and therefore have a strong prospect of future development. Notably, the currently available biosensor technologies for PAH monitoring have not so far been compiled together, and therefore a significant focus of this article is on biosensor technologies that are critical for timely detection and efficient management of PAHs. This review is focussed on inland aquatic ecosystems with an emphasis on fish biodiversity, as fish remains a major source of food and livelihood for a large proportion of the global population. This thought provoking study is likely to instigate new collaborative approaches for protecting aquatic biodiversity from PAHs-induced eco-toxicity.

Effects of supplementation diet containing Microcystis aeruginosa on haematological and biochemical changes in Labeo rohita infected with Aeromonas hydrophila.

An experiment of 100 days duration was conducted to test the effect of Microcystis aeruginosa in Labeo rohita against Aeromonas hydrophila. The rohu fingerlings (22 ± 2 g) were fed with the experimental diets incorporated with different concentrations of M. aeruginosa @ 0 g kg-1, 0.5 g kg-1, 1 g kg-1 and 5 g kg-1. Replicate groups of fish were fed for three month daily @4% body weight. At an interval of 30 days blood and serum samples were assayed for different haematological [total erythrocyte count (TEC), total leucocyte count (TLC), haemoglobin] and biochemical [blood glucose, serum aspartate aminotransferase (AST), serum alanine aminotransferase (ALT) and alkaline phosphatase (ALP)] parameters. Significantly (p ≤ 0.05) increased haemoglobin content and TLC was observed in Microcystis treated group. It was observed that the serum AST activity was significantly (p ≤ 0.05) decreased to all the treated groups of fish as compared to control on entire assay period. Serum ALT activity was significantly (p ≤ 0.05) different to all the treated groups of fish on day 60, on day 90 and on day 10 (except group B) bacterial post challenge as compared to control. After 90 days, fish were challenged with A. hydrophila and mortality (%) was recorded up to day 10 post challenge. Highest percentage of survival (72%) was noticed in the group fed 1.0 g Microcystis kg-1 dry diet. The present study suggests that the administration of bluegreen algae, Microcystis aeruginosa supplementation diets for 100 days protects the hematological and biochemical parameters in L. rohita from A. hydrophila.